CN102326677B - Composite micro-ecological feed additive and preparation method and premix thereof - Google Patents
Composite micro-ecological feed additive and preparation method and premix thereof Download PDFInfo
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Abstract
The invention discloses a composite micro-ecological feed additive and a premix thereof, and belongs to the technical field of feed science and feed additive. The feed additive contains multiple probiotics, and has the functions of enhancing the immunity of livestock and poultry and improving the production performance of the livestock and poultry; and meanwhile, the feed additive can be used as a feed mildew-proof agent, has good adhesion identifying and removing capacity on feed mycotoxins, can effectively inhibit the growth of mycete in the feed and adsorb and degrade the mycotoxins, is a novel, safe, reliable and pollution-free pure fermented multifunctional micro-ecological preparation product, and has good application prospect.
Description
Technical field
The present invention relates to a kind of composite microorganism type forage additives and premix thereof, belong to forage science and field of feed additive technology.
Background technology
Fast development along with aquaculture, people have related to various aspects to the research of feed addictive, and wherein the safety issue of antibiotic etc class feed addictive and feed have been become two large focuses of research in present feed industry and the animal husbandry production by mould and mycotoxin contamination problem.
Be subjected to the infringement of mould according to agricultural product such as the grain in the United Nations grain tissue estimation whole world annual nearly 5%～7%, feeds, thereby caused huge economic loss.By the feed of mould contamination, its nutriment is decomposed, and palatability descends, and produce multiple mycotoxin, animals eat the feed of moldy metamorphism, can occur acute or the slow poisoning symptom, cause the speed of growth of livestock and poultry slow, feed intake descends, digestibility reduces, fertility and immunity degradation, serious harm animal health, even cause and be poisoned to death, bring huge harm to aquaculture.
Mould inhibitor is of a great variety in the market, and common mould inhibitor has organic acid, acylate and organic acid ester and some composite mildew inhibitors.But these all are chemical anti mildew agents, and itself does not improve feed quality, in use animal are had excitant simultaneously, palatability, security and antibacterial aspect Shortcomings.
The feed probiotics is as a kind of novel Substitutes For Antibiotic, the pollution such as have no side effect because having, noresidue is polluted, do not develop immunity to drugs, have disease-resistant, cure the disease, the plurality of advantages such as diseases prevention, growth promotion, be subject to the welcome of aquaculture.At present, feeding micro-ecological preparation product on the market is more, action effect mainly concentrates on aspects such as improving animal immunizing power, increase animal production, both had the feedstuff mildew effect, has again the feed quality of improvement, improve the animal immunizing power effect, the product that increases animal production but rarely has report.
Summary of the invention
The object of the present invention is to provide a kind of raising immunity of livestock that has, have the new microbial forage additive of feedstuff mildew function concurrently.
Another object of the present invention provides the preparation method of this microbial forage additive and comprises the premix of this microbial forage additive.
The objective of the invention is to realize by following technical solution:
The invention provides a kind of composite microorganism type forage additives, this additive is a kind of pulvis, it is mainly mixed by the composition of following weight percentage: bacillus pumilus (Bacillus pumilus) bacterium powder 30-50%, lactobacillus acidophilus (Lactobacillus acidophilus) bacterium powder 20-30%, saccharomyces cerevisiae (saccharomyces cerevisiae) cell wall polysaccharides 0.5-1%, inulin 10%-30%, stone flour 1-10%.
Further, this additive is mainly mixed by the composition of following weight percentage: bacillus pumilus (Bacillus pumilus) bacterium powder 42%, lactobacillus acidophilus (Lactobacillus acidophilus) bacterium powder 28%, saccharomyces cerevisiae (saccharomyces cerevisiae) cell wall polysaccharides 1%, inulin 19%, stone flour 10%.
Wherein, bacillus pumilus bacterium powder of the present invention, lactobacillus acidophilus bacterium powder, brewing yeast cell wall polysaccharide can be selected disclosed bacterial strain preparation in the prior art, but for reaching best effects of the present invention, the present invention preferably uses deposit number to be the bacillus pumilus of CGMCC NO.4756, and deposit number is that the lactobacillus acidophilus of CGMCC No.5093 and the saccharomyces cerevisiae of the CGMCC No.2388 that deposit number is prepare bacterium powder or cell wall polysaccharides.
Wherein, described bacillus pumilus bacterium powder is dried under 40-90 ℃ of condition and is formed, and viable count is 5 * 10
8More than the CFU/g; Described lactobacillus acidophilus bacterium powder adopts the method for vacuum freeze drying, cryogenic temperature-55 ℃～-30 ℃, is prepared under vacuum 20～60Pa condition, and viable count is 2 * 10
7More than the CFU/g; In the described brewing yeast cell wall polysaccharide, gross protein value≤35%, beta glucan content 〉=30%, manna oligosacchride content 〉=20%; Inulin is food-grade raw materials; Stone flour is carrier.
Wherein, the concrete preparation method of composite microorganism type forage additives of the present invention is as follows:
The preparation of A, lactobacillus acidophilus bacterium powder: lactobacillus acidophilus is inoculated in the MRS culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 2 * 10
9CFU/ml; bacterium liquid by volume: protective agent is that 1: 10 proportioning adds protective agent (the protective agent prescription is skimmed milk 3%; trehalose 2%; manganese sulfate 0.5%, sucrose 0.2%, other is water); behind-80 ℃ of pre-freeze 4h; adopt the method for vacuum freeze drying, in cryogenic temperature-55～-30 ℃, make viable count greater than 2 * 10 under the vacuum 20-60Pa
7CFU/g lactobacillus acidophilus bacterium powder.
The preparation of B, bacillus pumilus bacterium powder: bacillus pumilus is inoculated in the LB culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 5 * 10
11CFU/ml, by weight bacterium liquid: carrier is that 1: 1 proportioning adds the carrier maize cob meal, and 60 ℃ of oven dry 24h make viable count greater than 5 * 10
8CFU/g bacillus pumilus bacterium powder.
The preparation of C, brewing yeast cell wall polysaccharide: the 1) preparation of S. cervisiae powder, the preparation method with reference to applicant's application number is: 200810106520.5 Chinese invention patent, get S. cervisiae powder 1000g, add 5L sodium hydroxide solution (1mol/L), at 100 ℃ of lower heating hydrolysis 4h, wait be cooled to room temperature after, 1000 * g, centrifugal 10min carries out Separation of Solid and Liquid; 2) after the abandoning supernatant, add 1L water in the residue insoluble matter, stir, so repeat twice centrifugal (1000 * g, 10min) again; 3) add in the insoluble matter 1.5L acetum (10%, V/V) stir, then in 85 ℃ of water-baths, be incubated 2h; 4) such as method 1) as described in method carry out Separation of Solid and Liquid, and washed twice; 5) after the residue insoluble matter washed three times with acetone, drying obtained the yeast cell wall polyose: gross protein value≤35%, beta glucan content 〉=30%, the brewing yeast cell wall polysaccharide of manna oligosacchride content 〉=20%.
D, take by weighing each composition raw material by proportioning, raw material is put into mixer, wherein, the preferred mixture homogeneity coefficient of variation of mixer mixes, and get final product less than 5% double-shaft paddle formula high efficient mixer.
The present invention also provides a kind of premix that comprises this composite microorganism type forage additives, and microbial forage additive adds percentage by weight in premix be 1-10 ‰.
Bacillus pumilus of the present invention (Bacillus pumilus) strain that is separated at home first for the applicant derives from the new bacillus pumilus of animal alimentary canal, from the cud of ox, separate a bacillus dominant strain that obtains for the applicant, through Physiology and biochemistry and 16s RNA Analysis and Identification, show that this Pseudomonas is in bacillus pumilus, its Classification And Nomenclature is bacillus pumilus (Bacillus pumilus), be preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on April 8th, 2011, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, be called for short CGMCC, deposit number is CGMCC No.4756.Saccharomyces cerevisiae among the present invention (Saccharomyces cerevisiae) has good stomach juice-resistant, bile tolerance, high temperature resistant, anti-adversity and the product acid such as tolerance common antibiotics, produce enzyme, suppress the living functions of benefit such as pathogen, for the applicant separates from healthy animal enteron aisle or ight soil, seed selection obtains, identify through Chinese agriculture microorganism fungus kind preservation administrative center, and being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on March 3rd, 2008, preserving number is that (the preservation proof is CGMCC No.2388: 200810106520.5 Chinese invention patent) referring to applicant's application number.Lactobacillus acidophilus of the present invention (Lactobacillus acidophilus) obtains for the applicant separates from animal intestinal, can secrete antibiotin class material (acidolin, bacillus acidophilus's element, lactein), antagonism to the pathogenic entero becteria generation, the propagation that suppresses the enteron aisle undesirable microorganism, adjust the gut flora balance, the applicant has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation on July 26th, 2011, preservation address and be called for short the samely, preserving number is CGMCCNo.5093.
Beneficial effect of the present invention is: the strain that used bacillus pumilus CGMCC NO.4756 is separated to first at home for the applicant among the present invention derives from the new bacillus pumilus of animal alimentary canal, and it is applied in the animal premix as feed addictive, first Application is in the Midew preventive for feed aspect simultaneously; And this bacterial strain has resistant to elevated temperatures characteristics, thereby has and do not need to be coated with the advantage that can directly granulate; Bacillus pumilus of the present invention is added in the premix as a kind of feed addictive, can improve feed quality, improve the content of crude protein in the feed, reduce coarse-fibred content in the feed, more conventional mould inhibitor does not contain chemical agent except safety, pollution-free, direct granulation simultaneously, and not only anti-mold effect is good, antibacterial power is strong, and can improve feed quality; Yeast cell wall polysaccharide among the present invention is to be prepared from through the alkali preparation method by S. cervisiae powder CGMCC No.2388, this product is rich in the biological immune factors such as beta glucan and functional oligosaccharides, can significantly strengthen cytophagous activity and phagocytic activity, strengthen the function of animal immune system, improve microbial environment in the animal body, identification adheres to and eliminates Mycotoxins in Feed, improves food conversion ratio.The used lactobacillus acidophilus CGMCC of the present invention No.5093 has stronger acid producing ability, the adding of lactobacillus acidophilus, not only can create slightly acidic environment, strengthen the stability of mould inhibitor, also can give feed good organoleptic properties, strengthening the animal intestinal resistance, prevention and treatment diarrhoea, promoting the feed safety aspect has good benefit to give birth to effect.
Probiotics of the present invention is by bacillus pumilus, lactobacillus acidophilus, yeast cell wall polysaccharide and inulin is composite forms, palatability is strong, not only has the reinforcement animal immune system, the selective animal intestinal beneficial bacterium propagation that promotes, suppress the field planting of beneficial bacterium and breed, improve microbial environment in the animal body, stimulate immune response, strengthen immunity of livestock, improve livestock and poultry production performance, reducing antibiotic uses, mycotoxins in feed there are good identification adhesion and elimination ability, the function of improving food conversion ratio; Can be used as again simultaneously a kind of Midew preventive for feed; It is few also to have addition, and preparation technology is simple, has no side effect, and the advantages such as noresidue have higher market value.
The specific embodiment
Embodiment 1: the preparation of composite microorganism type forage additives
The preparation of A, lactobacillus acidophilus bacterium powder: lactobacillus acidophilus CGMCC NO.5093 is inoculated in the MRS culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 2 * 10
9CFU/ml; by bacterium liquid: 1: 10 proportioning of protective agent (volume ratio) adds protective agent, and (the protective agent prescription is skimmed milk 3%; trehalose 2%; manganese sulfate 0.5%, sucrose 0.2%, other is water); behind-80 ℃ of pre-freeze 4h; adopt the method for vacuum freeze drying, in cryogenic temperature-50 ℃, making viable count under the vacuum 30Pa is 2 * 10
8CFU/g lactobacillus acidophilus bacterium powder.
The preparation of B, bacillus pumilus bacterium powder: bacillus pumilus CGMCC NO.4756 is inoculated in the LB culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 5 * 10
11CFU/ml, press bacterium liquid: 1: 1 proportioning of carrier (weight ratio) adds the carrier maize cob meal, 60 ℃ of oven dry 24h, making viable count is 5 * 10
9CFU/g bacillus pumilus bacterium powder.
The preparation of C, brewing yeast cell wall polysaccharide: 1) get S. cervisiae powder 1000g, add 5L sodium hydroxide solution (1mol/L), at 100 ℃ of lower heating hydrolysis 4h, wait be cooled to room temperature after, centrifugal (1000 * g, 10min) carry out Separation of Solid and Liquid; 2) after the abandoning supernatant, add 1L water in the residue insoluble matter, stir, so repeat twice centrifugal (1000 * g, 10min) again; 3) add in the insoluble matter 1.5L acetum (10%, V/V) stir, then in 85 ℃ of water-baths, be incubated 2h; 4) such as method 1) as described in method carry out Separation of Solid and Liquid, and washed twice; 5) after the residue insoluble matter washed three times with acetone, drying obtains the yeast cell wall polyose: gross protein value was 35%, and beta glucan content is 30%, and manna oligosacchride content is 20% brewing yeast cell wall polysaccharide.
D, take by weighing each composition raw material by proportioning, proportioning is as follows: bacillus pumilus bacterium powder 35%, lactobacillus acidophilus bacterium powder 30%, brewing yeast cell wall polysaccharide 0.5%, inulin 29.5%, stone flour 5%; Raw material is put into the mixture homogeneity coefficient of variation less than 5% double-shaft paddle formula high efficient mixer, mix, and get final product.
Embodiment 2: the preparation of composite microorganism type forage additives
The preparation of A, lactobacillus acidophilus bacterium powder: lactobacillus acidophilus CGMCC NO.5093 is inoculated in the MRS culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 2 * 10
9CFU/ml; by bacterium liquid: 1: 10 proportioning of protective agent (volume ratio) adds protective agent, and (the protective agent prescription is skimmed milk 3%; trehalose 2%; manganese sulfate 0.5%, sucrose 0.2%, other is water); behind-80 ℃ of pre-freeze 4h; adopt the method for vacuum freeze drying, in cryogenic temperature-50 ℃, make viable count 5 * 10 under the vacuum 30Pa
8CFU/g lactobacillus acidophilus bacterium powder.
The preparation of B, bacillus pumilus bacterium powder: bacillus pumilus CGMCC NO.4756 is inoculated in the LB culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 5 * 10
11CFU/ml, press bacterium liquid: 1: 1 proportioning of carrier (weight ratio) adds the carrier maize cob meal, and 60 ℃ of oven dry 24h make viable count greater than 8 * 10
9CFU/g bacillus pumilus bacterium powder.
The preparation of C, brewing yeast cell wall polysaccharide: 1) get S. cervisiae powder 1000g, add 5L sodium hydroxide solution (1mol/L), at 100 ℃ of lower heating hydrolysis 4h, wait be cooled to room temperature after, centrifugal (1000 * g, 10min) carry out Separation of Solid and Liquid; 2) after the abandoning supernatant, add 1L water in the residue insoluble matter, stir, so repeat twice centrifugal (1000 * g, 10min) again; 3) add in the insoluble matter 1.5L acetum (10%, V/V) stir, then in 85 ℃ of water-baths, be incubated 2h; 4) such as method 1) as described in method carry out Separation of Solid and Liquid, and washed twice; 5) after the residue insoluble matter washs three times with acetone, drying, obtaining yeast cell wall polyose gross protein value is 30%, and beta glucan content is 45%, and manna oligosacchride content is 30% brewing yeast cell wall polysaccharide.
D, take by weighing each composition raw material by proportioning, proportioning is as follows: bacillus pumilus bacterium powder 42%, lactobacillus acidophilus bacterium powder 28%, brewing yeast cell wall polysaccharide 1%, inulin 19%, stone flour 10%; Raw material is put into the mixture homogeneity coefficient of variation less than 5% double-shaft paddle formula high efficient mixer, mix, and get final product.
Embodiment 3: the preparation of composite microorganism type forage additives
The preparation of A, lactobacillus acidophilus bacterium powder: lactobacillus acidophilus CGMCC NO.5093 is inoculated in the MRS culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 2 * 10
9CFU/ml; by bacterium liquid: 1: 10 proportioning of protective agent (volume ratio) adds protective agent, and (the protective agent prescription is skimmed milk 3%; trehalose 2%; manganese sulfate 0.5%, sucrose 0.2%, other is water); behind-80 ℃ of pre-freeze 4h; adopt the method for vacuum freeze drying, in cryogenic temperature-50 ℃, making viable count under the vacuum 30Pa is 6 * 10
8CFU/g lactobacillus acidophilus bacterium powder.
The preparation of B, bacillus pumilus bacterium powder: bacillus pumilus CGMCC NO.4756 is inoculated in the LB culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 5 * 10
11CFU/ml, press bacterium liquid: 1: 1 proportioning of carrier (weight ratio) adds the carrier maize cob meal, and 60 ℃ of oven dry 24h make viable count greater than 4 * 10
9CFU/g bacillus pumilus bacterium powder.
The preparation of C, brewing yeast cell wall polysaccharide: 1) get S. cervisiae powder 1000g, add 5L sodium hydroxide solution (1mol/L), at 100 ℃ of lower heating hydrolysis 4h, wait be cooled to room temperature after, centrifugal (1000 * g, 10min) carry out Separation of Solid and Liquid; 2) after the abandoning supernatant, add 1L water in the residue insoluble matter, stir, so repeat twice centrifugal (1000 * g, 10min) again; 3) add in the insoluble matter 1.5L acetum (10%, V/V) stir, then in 85 ℃ of water-baths, be incubated 2h; 4) such as method 1) as described in method carry out Separation of Solid and Liquid, and washed twice; 5) after the residue insoluble matter washs three times with acetone, drying, obtaining yeast cell wall polyose gross protein value is 25%, and beta glucan content is 35%, and manna oligosacchride content is 31% brewing yeast cell wall polysaccharide.
D, take by weighing each composition raw material by proportioning, proportioning is as follows: bacillus pumilus bacterium powder 50%, lactobacillus acidophilus bacterium powder 22%, brewing yeast cell wall polysaccharide 1%, inulin 17%, stone flour 10%; Raw material is put into the mixture homogeneity coefficient of variation less than 5% double-shaft paddle formula high efficient mixer, mix, and get final product.
Embodiment 4, the application of composite microorganism type forage additives aspect the weanling pig cultivation
Selecting 144 average weights is the Da Bai * white two-way cross weanling pig of length of 8.65 ± 0.42kg, Weaning Age is 28 ± 2d, be divided at random 4 processing according to its body weight, sex, the not consistent principle of nest, the basal feed (blank group) of feeding respectively, add the feed addictive of embodiment 2 preparations in the basal feed, adding proportion is 0.5 ‰ (testing 1 group), 5 ‰ (testing 2 groups), the daily ration of 10 ‰ (testing 3 groups), 6 repetitions of every processing, each repeats 6 pigs, male and female half and half, and experimental period is 35d.Experimental result such as table 1.
The result shows, the equal feed consumption rate of experimental group average net gain in latter stage, daily gain and head that adds compound micro-ecological preparation all is higher than the blank group, this shows, the interpolation of this feeding micro-ecological preparation has the growth performance of Effective Raise weanling pig, reduce feedstuff-meat ratio, reduce the effect of the diarrhea rate of piglet.Analysis-by-synthesis draws, and the preferred addition of compound micro-ecological preparation is 5 ‰, and at this moment, experimental group is compared with the blank group, and piglet average net gain in latter stage improves 33.7%, and Average weight increasing a day improves 33.5%, and the material anharmonic ratio significantly is lower than control group, and the grice diarrhoea rate is 0.
Table 1 various dose probiotics is on the impact of Growth Performance of Weaning Piglets
Annotate: colleague's numeral indicates same letter person and represents difference not remarkable (p＞0.05), and different alphabetical persons represent significant difference (p＜0.05).
Embodiment 5: add compound micro-ecological preparation in the daily ration that goes mouldy to the impact of meat chick immunologic function
Select 450 of 7 age in days healthy AA meat chicks, be divided at random 5 groups by body weight, every group of 6 repetitions, each repeats 15 chickens.Grouping situation such as table 2.
Liver is the target organ of aflatoxin effect, but is not unique damaged organ.The meat chick is searched for food behind the aflatoxin, at first hepatic injury can occur, searches for food for a long time also to cause other organ to be subject in various degree damage.The corn that goes mouldy can significantly improve the relative weight of broiler chicken liver, heart, kidney, spleen.In addition, aflatoxin can bring out immunosupress, and Development of Immune Organs is obstructed, and causes the spleen enlargement, the bursa of farbricius, atrophy of thymus gland.
Table 2 experimental design
| Group | Daily ration forms |
| Control group | Normal corn |
| The moldy feed group | Normal corn in the daily ration is substituted by fermented maize |
| Test group I | The compound micro-ecological preparation of moldy feed+0.5 ‰ embodiment, 1 preparation |
| Test group II | The compound micro-ecological preparation of moldy feed+5 ‰ embodiment, 1 preparation |
| Test group III | The compound micro-ecological preparation of moldy feed+10 ‰ embodiment, 1 preparation |
Studies show that of this experiment, moldy feed group liver is obvious tumefaction when test 7d and 21d, and obvious enlargement also appears in spleen during 7d, and atrophy appears in thymus gland and the bursa of farbricius.Compare with the moldy feed group, the swelling of broiler chicken internal organs and the atrophy of immune organ phenomenon of test group have obtained alleviation, add 10 ‰ probiotics groups and have significantly improved the relative weight of the bursa of farbricius, and make it be higher than the control group level.Therefore, compound micro-ecological preparation of the present invention can reduce the feed that goes mouldy to a certain extent to the toxic action of internal organ, and has promoted the growth of immune organ.Shown in experimental result such as the table 3, table 4.
Table 3 compound micro-ecological preparation is on the impact (g/kg) of meat chick immune organ weight
Annotate: colleague's numeral indicates same letter person and represents difference not remarkable (p＞0.05), and different alphabetical persons represent significant difference (p＜0.05).
Table 4 compound micro-ecological preparation is on the impact (g/kg) of meat chick immune organ relative weight
Annotate: colleague's numeral indicates same letter person and represents difference not remarkable (p＞0.05), and different alphabetical persons represent significant difference (p＜0.05).
Embodiment 6: the application of compound micro-ecological preparation aspect the mildew-resistant of growing and fattening pigs complete diet pellet
Fresh pig complete diet pellet 30kg is divided into three groups, and control group does not add mould inhibitor, and test group I adds 50% calcium propionate Midew preventive for feed by 1 ‰ addition, and test group II adds the compound micro-ecological preparation of the embodiment of the invention 2 preparations by 5 ‰ addition.According to aspergillus flavus optimum growing condition in the bibliographical information, to test feed and leave 30 ℃ in, humidity is to preserve in 75% the incubator, every 10d gathers Feed Sample, test period is 60d, get 20g for each every group, detect aflatoxin in the feed with the ELISA method, experimental result is as shown in table 5.
Table 5 compound micro-ecological preparation is to the anti-mold effect of complete feed for fattening pigs
Studies show that, experiment 60d, control group feed aflatoxin content is up to 76.18 μ g/kg, and test group I feed aflatoxin content is 15.18 μ g/kg, aflatoxin content is 14.14 μ g/kg in the test group II feed, is lower than 20 μ g/kg of national forage health standard.Therefore, even in the optimum condition of aspergillus flavus growth and breeding, but add the generation of 5 ‰ compound micro-ecological preparation establishment Mycotoxins in Feed in feed, the calcium propionate class mould inhibitor effect of commonly using in its anti-mold effect and the feed is basic identical.
Claims (5)
1. composite microorganism type forage additives, it is characterized in that: it is a kind of pulvis, this additive is mainly mixed by the composition of following weight percentage: bacillus pumilus (Bacillus pumilus) bacterium powder 30-50%, lactobacillus acidophilus (Lactobacillus acidophilus) bacterium powder 20-30%, saccharomyces cerevisiae (saccharomyces cerevisiae) cell wall polysaccharides 0.5-1%, inulin 10%-30%, stone flour 1-10%, the deposit number of described bacillus pumilus is CGMCC NO.4756, and the deposit number of described lactobacillus acidophilus is CGMCC No.5093; The deposit number of described saccharomyces cerevisiae is CGMCC No.2388.
2. composite microorganism type forage additives as claimed in claim 1, it is characterized in that: this additive is mainly mixed by the composition of following weight percentage: bacillus pumilus (Bacillus pumilus) bacterium powder 42%, lactobacillus acidophilus (Lactobacillus acidophilus) bacterium powder 28%, saccharomyces cerevisiae (saccharomyces cerevisiae) cell wall polysaccharides 1%, inulin 19%, stone flour 10%.
3. composite microorganism type forage additives as claimed in claim 1, it is characterized in that: described bacillus pumilus bacterium powder is dried under 40-90 ℃ of condition and is formed, and viable count is 5 * 10
8More than the CFU/g; Described lactobacillus acidophilus bacterium powder adopts the method for vacuum freeze drying, cryogenic temperature-55 ℃～-30 ℃, is prepared under vacuum 20～60Pa condition, and viable count is 2 * 10
7More than the CFU/g; In the described brewing yeast cell wall polysaccharide, gross protein value≤35%, beta glucan content 〉=30%, manna oligosacchride content 〉=20%.
4. the preparation method of each described composite microorganism type forage additives of claim 1-3 is characterized in that: comprise the steps:
The preparation of A, lactobacillus acidophilus bacterium powder: lactobacillus acidophilus is inoculated in the MRS culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 2 * 10
9CFU/ml, by volume bacterium liquid: protective agent is that 1: 10 proportioning adds protective agent, and the protective agent prescription is as follows: skimmed milk 3%, and trehalose 2%, manganese sulfate 0.5%, sucrose 0.2%, other is water; Adopt the method for vacuum freeze drying, in cryogenic temperature-55～-30 ℃, make viable count greater than 2 * 10 under the vacuum 20-60Pa
7CFU/g lactobacillus acidophilus bacterium powder;
The preparation of B, bacillus pumilus bacterium powder: bacillus pumilus is inoculated in the LB culture medium, and 37 ℃ of static cultivation 12h adjust bacteria concentration to 5 * 10
11CFU/ml, by weight bacterium liquid: carrier is that 1: 10 proportioning adds the carrier maize cob meal, and 60 ℃ of oven dry 24h make viable count greater than 5 * 10
8CFU/g bacillus pumilus bacterium powder;
The preparation of C, brewing yeast cell wall polysaccharide: 1) preparation S. cervisiae powder, get S. cervisiae powder 1000g, adding 5L concentration is the sodium hydroxide solution of 1mol/L, at 100 ℃ of lower heating hydrolysis 4h, wait be cooled to room temperature after, 1000 * g, centrifugal 10min carries out Separation of Solid and Liquid; 2) after the abandoning supernatant, add 1L water in the residue insoluble matter, stir, centrifugal 10min under 1000 * g repeats twice; 3) adding the 1.5L volume ratio in the insoluble matter is 10% acetum, stirs, and then is incubated 2h in 85 ℃ of water-baths; 4) such as step 1) as described in method carry out Separation of Solid and Liquid, and washed twice; 5) after the residue insoluble matter washed three times with acetone, drying obtained the yeast cell wall polyose: gross protein value≤35%, beta glucan content 〉=30%, the brewing yeast cell wall polysaccharide of manna oligosacchride content 〉=20%;
D, take by weighing each composition raw material by proportioning, raw material is put into mixer, mix, and get final product.
5. premix that comprises each described composite microorganism type forage additives of claim 1-3 is characterized in that: the percentage by weight that composite microorganism type forage additives adds in premix is 1-10 ‰.
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| CN102559539B (en) * | 2011-12-02 | 2015-04-15 | 北京大北农科技集团股份有限公司 | Lactobacillus acidophilus and application thereof and feed additive thereof and premix compound thereof |
| CN103243028B (en) * | 2012-02-02 | 2015-11-25 | 安琪酵母股份有限公司 | Yeast cells wall and method for making, fodder additives and method for making thereof and feed |
| CN102885206B (en) * | 2012-10-29 | 2013-10-30 | 曾伟 | Natural animal feed additive |
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| CN101978849B (en) * | 2008-05-14 | 2012-10-03 | 漳州大北农农牧科技有限公司 | Composite micro-ecological preparation and application thereof |
| CN102160595B (en) * | 2011-02-22 | 2012-12-12 | 福建省新闽科生物科技开发有限公司 | Preparation process of complex microorganism fermented active feed |
| CN102204632B (en) * | 2011-05-12 | 2013-03-27 | 成都和谐生物科技有限公司 | Microbial feed additive for increasing livestock and poultry product quality and improving meat egg milk flavor and preparation method thereof |
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Application publication date: 20120125 Assignee: JIANGXI GAOAN DABEINONG FORAGE CO., LTD. Assignor: Zhangzhou agricultural farming and animal husbandry science and Technology Co., Ltd.|Beijing DaBeiNong Technology Group Limited by Share Ltd|Inner Mongolia sijichun Feed Co. Ltd. Contract record no.: 2015990000032 Denomination of invention: Composite micro-ecological feed additive and preparation method and premix thereof Granted publication date: 20131016 License type: Exclusive License Record date: 20150119 |
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Assignee: JIANGXI GAOAN DABEINONG FORAGE CO., LTD. Assignor: Zhangzhou agricultural farming and animal husbandry science and Technology Co., Ltd. |Beijing DaBeiNong Technology Group Limited by Share Ltd| Inner Mongolia sijichun Feed Co. Ltd. Contract record no.: 2015990000032 Date of cancellation: 20160115 |
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Effective date of registration: 20160429 Address after: 100080 Beijing City, Haidian District Zhongguancun street, No. 27 Zhongguancun building 14 DaBeiNong group Patentee after: Beijing Dabeinong Technology Group Co., Ltd. Patentee after: JIANGXI GAOAN DABEINONG FORAGE CO., LTD. Address before: 100080 Beijing City, Haidian District Zhongguancun street, No. 27 Zhongguancun building 14 DaBeiNong group Patentee before: Beijing Dabeinong Technology Group Co., Ltd. Patentee before: Inner Mongolia Ever Spring Feed Co., Ltd. Patentee before: Zhangzhou Dabeinong Agriculture & Pasture Technology Co., Ltd. Patentee before: JIANGXI GAOAN DABEINONG FORAGE CO., LTD. |